title: Selection of recombinant antibodies by phage display technology and application for detection of allergenic Brazil nut (Bertholletia excelsa) in processed foods.
abstract: Current immunological methods for detection of Brazil nut allergens in foods are based on polyclonal antibodies raised in animals. Phage display technology allows the procurement of high-affinity antibodies avoiding animal immunization steps and therefore attaining the principle of replacement supported by animal welfare guidelines. In this study, we screened Tomlinson I and J libraries for specific binders against Brazil nut by employing a Brazil nut protein extract and a purified Brazil nut 2S globulin, and we successfully isolated a phage single chain variable fragment (named BE95) that specifically recognizes Brazil nut proteins. The selected phage scFv was further used as affinity probe to develop an indirect phage-ELISA for detection of Brazil nut in experimental binary mixtures and in commercial food products, with a limit of detection of 5 mg g(-1). This study describes for the first time the isolation of recombinant antibody fragments specific for an allergenic tree nut protein from a naïve library and paves the way to develop new immunoassays for food analysis based on probes that can be produced in vitro when required and do not rely on animal immunization.
authors.affiliation: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos Facultad de Veterinaria, Universidad Complutense de Madrid 28040 Madrid, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: de la Cruz
authors.forename: Inés María
authors.initials: IM
authors.lastname: López-Calleja
authors.forename: Marcos
authors.initials: M
authors.lastname: Alcocer
authors.forename: Isabel
authors.initials: I
authors.lastname: González
authors.forename: Rosario
authors.initials: R
authors.lastname: Martín
authors.forename: Teresa
authors.initials: T
authors.lastname: García
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D031383:Bertholletia / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001938:Brazil; D061505:Cell Surface Display Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D015723:Gene Library; D006801:Humans; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology
chemical_list: D000485:Allergens; D010940:Plant Proteins; D057127:Single-Chain Antibodies